Water stable iii-v semiconductor nanocrystal complexes and methods of making same

ABSTRACT

A water-stable semiconductor nanocrystal complex that is stable and has high luminescent quantum yield. The water-stable semiconductor nanocrystal complex has a semiconductor nanocrystal core of a III-V semiconductor nanocrystal material and a water-stabilizing layer. A method of making a water-stable semiconductor nanocrystal complex is also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation of U.S. application Ser. No.11/353,957, filed on Feb. 15, 2006, which is a continuation-in-part ofU.S. application Ser. No. 11/125,129, filed on May 10, 2005, whichclaims priority to U.S. Provisional Application No. 60/569,452, filedMay 10, 2004. All applications are incorporated by reference herein.

FIELD OF THE INVENTION

The present invention relates to water-stable semiconductor nanocrystalcomplexes and particularly to water-stable III-V semiconductornanocrystal complexes. The present invention also relates to methods ofmaking water-stable semiconductor nanocrystal complexes.

BACKGROUND OF THE INVENTION

Semiconductor nanocrystals are typically tiny crystals of II-VI, III-V,IV-VI materials that have a diameter between 1 nanometer (nm) and 20 nm.In the strong confinement limit, the physical diameter of thenanocrystal is smaller than the bulk excitation Bohr radius causingquantum confinement effects to predominate. In this regime, thenanocrystal is a 0-dimensional system that has both quantized densityand energy of electronic states where the actual energy and energydifferences between electronic states are a function of both thenanocrystal composition and physical size. Larger nanocrystals have moreclosely spaced energy states and smaller nanocrystals have the reverse.Because interaction of light and matter is determined by the density andenergy of electronic states, many of the optical and electric propertiesof nanocrystals can be tuned or altered simply by changing thenanocrystal geometry (i.e. physical size).

Single nanocrystals or monodisperse populations of nanocrystals exhibitunique optical properties that are size tunable. Both the onset ofabsorption and the photoluminescent wavelength are a function ofnanocrystal size and composition. The nanocrystals will absorb allwavelengths shorter than the absorption onset, however,photoluminescence will always occur at the absorption onset. Thebandwidth of the photoluminescent spectra is due to both homogeneous andinhomogeneous broadening mechanisms. Homogeneous mechanisms includetemperature dependent Doppler broadening and broadening due to theHeisenburg uncertainty principle, while inhomogeneous broadening is dueto the size distribution of the nanocrystals. The narrower the sizedistribution of the nanocrystals, the narrower the full-width half max(FWHM) of the resultant photoluminescent spectra. In 1991, Brus wrote apaper reviewing the theoretical and experimental research conducted oncolloidally grown semiconductor nanocrystals, such as cadmium selenide(CdSe) in particular. Brus L., Quantum Crystallites and NonlinearOptics, Applied Physics A, 53 (1991)). That research, precipitated inthe early 1980's by the likes of Efros, Ekimov, and Brus himself,greatly accelerated by the end of the 1980's as demonstrated by theincrease in the number of papers concerning colloidally grownsemiconductor nanocrystals.

Quantum yield (i.e. the percent of absorbed photons that are reemittedas photons) is influenced largely by the surface quality of thenanocrystal. Photoexcited charge carriers will emit light upon directrecombination but will give up the excitation energy as heat if photonor defect mediated recombination paths are prevalent. Because thenanocrystal may have a large surface area to volume ratio, dislocationspresent on the surface or adsorbed surface molecules having asignificant potential difference from the nanocrystal itself will tendto trap excited state carriers and prevent radioactive recombination andthus reduce quantum yield. It has been shown that quantum yield can beincreased by removing surface defects and separating adsorbed surfacemolecules from the nanocrystal by adding a shell of a semiconductor witha wider bulk bandgap than that of the core semiconductor.

Inorganic colloids have been studied for over a century ever sinceMichael Faraday's production of gold sols in 1857. Rossetti and Brusbegan work on semiconductor colloids in 1982 by preparing and studyingthe luminescent properties of colloids consisting of II-VIsemiconductors, namely cadmium sulfide (CdS). (Rossetti, R.; Brus L.,Electron-Hole Recombination Emission as a Probe of Surface Chemistry inAqueous CdS Colloids, J. Phys. Chem., 86, 172 (1982)). In that paper,they describe the preparation and resultant optical properties of CdScolloids, where the mean diameter of the suspended particles is greaterthan 20 nm. Because the sizes of the particles were greater than theexaction Bohr radius, quantum confinement effects that result in theblue shifting of the fluorescence peak was not observed. However,fluorescence at the bulk bandedge energies were observed and had a FWHMof 50-60 nm.

CdS colloids exhibiting quantum confinement effects (blue shifted maximain the absorption spectra) were being prepared since 1984. (Fotjik A.,Henglein A., Ber. Bunsenges. Phys. Chem., 88, (1984); Fischer C., FotjikA., Henglein A., Ber. Bunsenges. Phys. Chem., (1986)). In 1987, Spanheland Henglein prepared CdS colloids having mean particle diametersbetween 4 and 6 nm. (Spanhel L., Henglein A., Photochemistry ofColloidal Semiconductors, Surface Modification and Stability of StrongLuminescing CdS Particles, Am. Chem. Soc., 109 (1987)). The colloidsdemonstrated quantum confinement effects including the observation ofsize dependent absorption maxima (first exciton peaks) as well as sizedependent fluorescent spectra. The colloids were prepared by bubbling asulphur containing gas (H₂S) through an alkaline solution containingdissolved cadmium ions. The size and resultant color (of thefluorescence) of the resultant nanocrystals were dependent upon the pHof the solution. The colloids were further modified or “activated” bythe addition of cadmium hydroxide to the solution that coated thesuspended nanocrystals. The resultant core-shell nanocrystalsdemonstrated that the quantum yield of the photoluminescence wasincreased from under 1% to well over 50% with a FWHM of thephotoluminescent spectra under 50 nm for some of the preparations.

Kortan and Brus developed a method for creating CdSe coated zincsulphide (ZnS) nanocrystals and the opposite, zinc sulphide coatedcadmium selenide nanocrystals. (Kortan R., Brus L., Nucleation andGrowth of CdSe on ZnS Quantum Crystallite Seeds, and Vice Versa, inInverse Micelle Media, J. Am. Chem. Soc., 112 (1990)). The preparationgrew ZnS on CdSe “seeds” using a organometallic precursor-based reversemicelle technique and kept them in solution via an organic capping layer(thiol phenol). The CdSe core nanocrystals had diameters between 3.5 and4 nm and demonstrated quantum confinement effects including observableexciton absorption peaks and blue shifted photoluminescence. Usinganother preparation, CdSe cores were coated by a 0.4 nm layer of ZnS.The photoluminescence spectra of the resultant core-shell nanocrystalsindicates a peak fluorescence at 530 nm with an approximate 40-45 nmFWHM.

Murray and Bawendi developed an organometallic preparation capable ofmaking CdSe, CdS, and CdTe nanocrystals. (Murray C., Norris D., BawendiM., Synthesis and Characterization of Nearly Monodisperse CdE (E=S, Se,Te) Semiconductor Nanocrystallites, J. Am. Chem. Soc., 115, (1993)).This work, based on the earlier works of Brus, Henglein, Peyghambarian,allowed for the growth of nanocrystals having a diameter between 1.2 nmand 11.5 nm and with a narrow size distribution (<5%). The synthesisinvolved a homogeneous nucleation step followed by a growth step. Thenucleation step is initiated by the injection of an organometalliccadmium precursor (dimethyl cadmium) with a selenium precursor(TOPSe-TriOctylPhosphine Selenium) into a heated bath containingcoordinating ligands (TOPO-TriOctylPhosphineOxide). The precursorsdisassociate in the solvent, causing the cadmium and selenium to combineto form a growing nanocrystal. The TOPO coordinates with the nanocrystalto moderate and control the growth. The resultant nanocrystal solutionshowed an approximate 10% size distribution, however, by titrating thesolution with methanol the larger nanocrystals could be selectivelyprecipitated from the solution thereby reducing the overall sizedistribution. After size selective precipitation, the resultantnanocrystals in solution were monodisperse (capable of reaching a 5%size distribution) but were slightly prolate (i.e. nonspherical havingan aspect ratio between 1.1 and 1.3). The photoluminescence spectra showa FWHM of approximately 30-35 nm and a quantum yield of approximately9.6%.

Katari and Alivisatos slightly modified the Murray preparation to makeCdSe nanocrystals. (Katari J., Alivisatos A., X-ray PhotoelectronSpectroscopy of CdSe Nanocrystals with Applications to Studies of theNanocrystal Surface, J. Phys. Chem., 98 (1994)). They found that bysubstituting the selenium precursor TOPSe with TBPSe(TriButylPhosphineSelenide), nanocrystals were produced that weremonodisperse without size selective precipitation, were crystalline, andspherical. The nanocrystals were size tunable from 1.8 nm to 6.7 nm indiameter and had an exciton peak position ranging from 1.9-2.5 eV(corresponding to 635-496 nm wavelength). Like the Murray paper, TOPOwas used as the coordinating ligand.

Hines and Guyot-Sionest developed a method for synthesizing a ZnS shellaround a CdSe core nanocrystal. (Hines et al., “Synthesis andCharacterization of strongly Luminescing ZnS capped CdSe Nanocrystals”;J. Phys. Chem., 100:468-471 (1996)). The CdSe cores, having amonodisperse distribution between 2.7 nm and 3.0 nm (i.e. 5% sizedistribution with average nanocrystal diameter being 2.85 nm), wereproduced using the Katari and Alivisatos variation of the Murraysynthesis. The photoluminescence spectra of the core shows a FWHM ofapproximately 30 nm with a peak at approximately 540 nm. The core CdSenanocrystals were separated, purified, and resuspended in a TOPOsolvent. The solution was heated and injected with zinc and sulphurprecursors (dimethyl zinc and (TMS)₂S) to form a ZnS shell around theCdSe cores. The resultant shells were 0.6±0.3 nm thick, corresponding to1-3 monolayers. The photoluminescence of the core-shell nanocrystals hada peak at 545 nm, FWHM of 40 nm, and a quantum yield of 50%.

One exemplary problem associated with known colloidal shell synthesismethods is that they are limited in regard to the growth of high qualityIII-V semiconductor nanocrystals due, at least in part, to the covalentnature of the material. Past attempts at growing III-V semiconductornanocrystals have resulted in quantum yields of approximately 10%. Micicet al., J. Phys. Chem. B. 2000, 104, 12149-12156. It has been shownusing a slow growth technique that indium phosphate (InP) corenanocrystals with a shell of zinc sulphide (ZnS) may result in quantumyields of 20% but this process takes days to complete. Haubold et al.,ChemPhysChem 2001, 5, 331. Many applications of nanocrystal complexes,such as LEDs, inks, and pigments, require higher luminescent quantumyields.

Additionally, the growth of III-V nanocrystals is difficult because ofthe need for vacuum growth and strict humidity conditions. Etching hasshown to improve the quantum yield of III-V materials however, theetching process introduces problems when bonding ligands to the surfaceof the nanocrystals. This causes poor colloidal stability and makes thecomplexes incompatible for a variety of applications. In addition, theetched surface does not provide sufficient electronic passivation. As aresult, electronic interaction between a matrix material and thenanocrystal may result in fluorescence quenching.

Additionally, etching relies on photoexcitation, which requires dilutemixtures. This is not conducive to the scale up and production of largequantities of III-V semiconductor nanocrystal complexes. By readilylosing their fluorescence, these quantum dot materials are simplyunsuitable for most applications. Thus, it is necessary to develop astable III-V semiconductor nanocrystal complex that is brightlyfluorescing, and soluble in water.

SUMMARY OF THE INVENTION

In an embodiment, the present invention provides a water-stablesemiconductor nanocrystal complex. The water-stable semiconductornanocrystal complex comprises a semiconductor nanocrystal compositionincluding a III-V semiconductor nanocrystal core. The water-stablesemiconductor nanocrystal complex also includes a surface layercomprising molecules having a moiety with an affinity for thesemiconductor nanocrystal composition and a moiety with an affinity fora hydrophobic solvent. The water-stable semiconductor nanocrystalcomplex further comprises a water-stablizing layer having a hydrophobicend for interacting with the surface layer and a hydrophilic end forinteracting with an aqueous medium. The water-stable semiconductornanocrystal complex is electronically and chemically stable with aluminescent quantum yield of at least 25%.

Additionally, the present invention provides for a method of making awater stable semiconductor nanocrystal complex comprising synthesizing asemiconductor nanocrystal core comprising a semiconductor nanocrystalmaterial. The method further comprises forming a surface layer eitherdirectly or indirectly on the semiconductor nanocrystal core, andforming a water-stabilizing layer on the surface layer. Thewater-stabilizing layer has a hydrophobic end for interacting with thesurface layer and a hydrophilic end for interacting with an aqueousmedium.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic illustration of a semiconductor nanocrystalcomposition according to an embodiment of the present invention.

FIG. 2 is a schematic illustration of a semiconductor nanocrystalcomposition according to another embodiment of the present invention.

FIG. 3 is a schematic illustration of a water-stable semiconductornanocrystal complex according to an embodiment of the present invention.

FIG. 4 is an exemplary conjugation method to conjugate tertiarymolecules to a water-stable semiconductor nanocrystal complex of thepresent invention.

FIG. 5 is a flow chart illustrating a method of making a water-stablesemiconductor nanocrystal complex according to an embodiment of thepresent invention.

DETAILED DESCRIPTION OF THE INVENTION

Referring to FIG. 1, in an embodiment, the present invention provides asemiconductor nanocrystal composition 70 comprising a semiconductornanocrystal core 10 (also known as a semiconductor nanoparticle orsemiconductor quantum dot) having an outer surface 15. Semiconductornanocrystal core 10 may be spherical nanoscale crystalline materials(although oblate and oblique spheroids can be grown as well as rods andother shapes) having a diameter of less than the Bohr radius for a givenmaterial and typically but not exclusively comprises II-IV, III-V, orIV-VI binary semiconductors. Non-limiting examples of semiconductormaterials that semiconductor nanocrystal core 10 can comprise includeZnS, ZnSe, ZnTe, CdS, CdSe, CdTe, HgS, HgSe, HgTe (II-VI materials),PbS, PbSe, PbTe (IV-VI materials), AlN, AlP, AlAs, AlSb, GaN, GaP, GaAs,GaSb, InN, InP, InAs, InSb (III-V materials). In a preferred embodiment,semiconductor nanocrystal core 10 comprises III-V semiconductornanocrystal materials. In addition to binary semiconductors,semiconductor nanocrystal core 10 may comprise ternary semiconductormaterials. Non-limiting examples of ternary semiconductor materialsinclude AxByC wherein A may comprise a group II, III or IV element, Bmay comprise a group II, III, or IV element, and C may comprise a groupV or VI element, and x and y are molar fractions between 0 and 1.

In an embodiment, one or more metals 20 are formed on outer surface 15of semiconductor nanocrystal core 10 (referred to herein as “metallayer” 20). Metal layer 20 may act to passivate outer surface 15 ofsemiconductor nanocrystal core 10 and limit the diffusion rate of oxygenmolecules to semiconductor nanocrystal core 10. According to the presentinvention, metal layer 20 is formed on outer surface 15 after synthesisof semiconductor nanocrystal core 10 (as opposed to being formed onouter surface 15 concurrently during synthesis of semiconductornanocrystal core 10). Metal layer 20 may include any number, type,combination, and arrangement of metals. For example, metal layer 20 maybe simply a monolayer of metals formed on outer surface 15 or multiplelayers of metals formed on outer surface 15. Metal layer 20 may alsoinclude different types of metals arranged, for example, in alternatingfashion. Further, metal layer 20 may encapsulate semiconductornanocrystal core 10 as shown in FIG. 1 or may be formed on only parts ofouter surface 15 of semiconductor nanocrystal core 10. Metal layer 20may include the metal from which the semiconductor nanocrystal core ismade either alone or in addition to another metal. Non-limiting examplesof metals that may be used as part of metal layer 20 include Cd, Zn, Hg,Pb, Al, Ga, or In.

Semiconductor nanocrystal composition 70, according to the presentinvention, is electronically and chemically stable with a highluminescent quantum yield. Chemical stability refers to the ability of asemiconductor nanocrystal composition to resist fluorescence quenchingover time in aqueous and ambient conditions. Preferably, thesemiconductor nanocrystal compositions resist fluorescence quenching forat least a week, more preferably for at least a month, even morepreferably for at least six months, and most preferably for at least ayear. Electronic stability refers to whether the addition of electron orhole withdrawing ligands substantially quenches the fluorescence of thesemiconductor nanocrystal composition. Preferably, a semiconductornanocrystal composition would also be colloidally stable in that whensuspended in organic or aqueous media (depending on the ligands) theyremain soluble over time. A high luminescent quantum yield refers to aquantum yield of at least 25%. Quantum yield may be measured bycomparison to Rhodamine 6G dye with a 488 excitation source. Preferably,the quantum yield of the semiconductor nanocrystal composition is atleast 30%, more preferably at least 45%, and even more preferably atleast 55%, as measured under ambient conditions. The semiconductornanocrystal compositions of the present invention experience little lossof fluorescence over time and can be manipulated to be soluble inorganic and inorganic solvents as traditional semiconductornanocrystals.

Semiconductor nanocrystal core 10 and metal layer 20 may be grown by thepyrolysis of organometallic precursors in a chelating ligand solution orby an exchange reaction using the prerequisite salts in a chelatingligand solution. The chelating ligands are typically lyophilic and havean affinity moiety for the metal layer and another moiety with anaffinity toward the solvent, which is usually hydrophobic. Typicalexamples of chelating ligands include lyophilic surfactant moleculessuch as Trioctylphosphine oxide (TOPO), Trioctylphosphine (TOP), andTributylphosphine (TBP).

Referring to FIG. 2, in an alternative embodiment, the present inventionprovides a nanocrystal composition 70 further comprising a shell 150overcoating metal layer 20. Shell 150 may comprise a semiconductormaterial having a bulk bandgap greater than that of semiconductornanocrystal core 10. In such an embodiment, metal layer 20 may act topassivate outer surface 15 of semiconductor nanocrystal core 10 as wellas to prevent or decrease lattice mismatch between semiconductornanocrystal core 10 and shell 150.

Shell 150 may be grown around metal layer 20 and is typically between0.1 nm and 10 nm thick. Shell 150 may provide for a type A semiconductornanocrystal composition 70. Shell 150 may comprise various differentsemiconductor materials such as, for example, CdSe, CdS, CdTe, ZnS,ZnSe, ZnTe, HgS, HgSe, HgTe, InP, InAs, InSb, InN, GaN, GaP, GaAs, GaSb,PbSe, PbS, or PbTe.

One example of shell 150 that may be used to passivate outer surface 15of semiconductor nanocrystal core 10 is ZnS. The presence of metal layer20 may provide for a more complete and uniform shell 150 without theamount of defects that would be present with a greater lattice mismatch.Such a result may improve the quantum yield of resulting nanocrystalcomposition 70.

Semiconductor nanocrystal core 10, metal layer 20, and shell 150 may begrown by the pyrolysis of organometallic precursors in a chelatingligand solution or by an exchange reaction using the prerequisite saltsin a chelating ligand solution. The chelating ligands are typicallylyophilic and have an affinity moiety for the metal layer and anothermoiety with an affinity toward the solvent, which is usuallyhydrophobic. Typical examples of chelating ligands include lyophilicsurfactant molecules such as Trioctylphosphine oxide (TOPO),Trioctylphosphine (TOP), and Tributylphosphine (TBP).

Referring to FIG. 3, the present invention also provides a water-stablesemiconductor nanocrystal complex 100 comprising a semiconductornanocrystal composition described above and further comprising a surfacelayer 50 that coats the semiconductor nanocrystal composition. Surfacelayer 50 generally comprises surface organic molecules that have amoiety 51 with an affinity for semiconductor nanocrystal composition 70and another moiety 52 with an affinity for a hydrophobic solvent.Moieties that have an affinity for a nanocrystal surface include thiols,amines, phosphines, and phosphine oxides. Non-limiting examples ofsurface molecules comprising surface layer 50 include trioctyl phosphineoxide (TOPO), trioctyl phosphine (TOP), tributyl phosphine (TBP),dodecyl amine, octadecyl amine, hexadecylamine, stearic acid, oleicacid, palmitic acid, lauric acid, and any combinations thereof. Suchsurface molecules are typically used in the synthesis of semiconductornanocrystal compositions and can remain on the surface of thenanocrystals after synthesis or may be replaced by other surfactantsafter synthesis. As is generally known to one of skill in the art,semiconductor nanocrystals according to the present invention may becoated with a surface layer by pyrolysis of organometallic percursors ina chelating ligand solution or by an exchange reaction using theprerequisite salts in a chelating surface solution, such chelatingsurfaces typically being lyophilic.

Referring still to FIG. 3, a water-stable semiconductor nanocrystalcomplex of the present invention further comprises a water-stabilizinglayer 60. Water stabilizing layer 60 preferably comprises lipids orpolymer-based small molecules including amino acids with a hydrophilicend section 62 and a hydrophobic end section 61. The hydrophobic endsection of a water-stabilizing layer has an affinity for a surface layerpresent on the surface of a semiconductor nanocrystal composition.

In an embodiment, a water-stabilizing layer comprises diblock polymersthat surround the semiconductor nanocrystal composition to form amicelle. More than one surface-coated semiconductor nanocrystalcomposition of the present invention may be surrounded by a diblockpolymer coating. A diblock polymer is generally but not exclusively alinear chain that has a hydrophobic end comprising hydrophobicfunctional groups that is covalently bonded to a hydrophilic endcomprising hydrophilic functional groups. In an aqueous medium, adiblock polymer coating assembles around a surface-coated semiconductornanocrystal composition of the present invention. Specifically, thehydrophobic end of a diblock polymer is attracted to the surface-coatednanocrystal composition and interacts with the moiety of the surfacelayer that has an affinity for a hydrophobic solvent through noncovalentinteractions such as, for example, hydrogen bonding, Van der WaalsForces, and hydrophobic interactions. The hydrophilic end of a diblockpolymer, in turn, is directed to the aqueous medium.

With respect to the lengths of the hydrophobic and hydrophilic end of adiblock polymer, each end has lengths greater than 1 and preferably eachhave lengths between 1 and 1000. In a more preferred embodiment, thehydrophobic end of a diblock polymer has between 60 and 180 carbonatoms. In a more preferred embodiment, the hydrophobic end has about 150carbon atoms and the hydrophilic end has about 220-240 carbon atoms.Also in a preferred embodiment, the hydrophobic end has about 10-20monomer units and the hydrophilic end has about 110-120 monomer units.Although the hydrophilic end and the hydrophobic end may have differentlengths, in a preferred embodiment, they are substantially equal inlength.

The hydrophobic functional groups of the hydrophobic end of a diblockpolymer that can be used as a water-stabilizing layer are preferablygroups of covalently bonded atoms on a larger molecule that are nonpolarand not ionizable and therefore have an affinity for nonpolar andnon-ionizable solvents. Non-limiting examples of hydrophobic functionalgroups according to the present invention include hydrocarbons ofvarious lengths. The hydrophilic functional groups of the hydrophilicend are preferably groups of atoms on a larger molecule that are highlypolar or ionizable and therefore have an affinity for water and otherpolar solvents. Non-limiting examples of hydrophilic functional groupsinclude hydroxy, amine, carboxyl, sulfonates, phosphates, amines,nitrates, and any combinations thereof.

Non-limiting examples of diblock polymers that comprise a diblockpolymer coating surrounding a surface-coated semiconductor nanocrystalcomposition according to an embodiment of the present invention includepoly(acrylic acid-b-methyl methacrylate), poly(methylmethacrylate-b-sodium acrylate), poly(t-butyl methacrylate-b-ethyleneoxide), poly(methyl methacrylate-b-sodium methacrylate), poly(methylmethacrylate-b-N-methyl 1-4vinyl pyridinium iodide), poly(methylmethacrylate-b-N,N-dimethyl acrylamide), poly(butadiene-b-methacrylateacid and sodium salt), poly(butadiene(1,2 addition)-b-acrylic acid),poly(butadiene(1,2 addition)-b-sodium acrylate), poly(butadiene(1,4addition)-b-acrylic acid), poly(butadiene(1,4 addition)-b-sodiumacrylate), poly(butadiene(1,4 addition)-b-ethylene oxide),poly(butadiene(1,2 addition)-b-ethylene oxide), poly(styrene-b-acrylicacid), poly(styrene-b-acrylamide), poly(styrene-b-cesium acrylate),poly(styrene-b-sodium acrylate), poly(styrene-b-ethylene oxide),poly(styrene-b-methacrylic acid), poly(styrene-b-sodium methacrylate),and any combinations thereof.

In order to form a cohesive coating around a surface-coatedsemiconductor nanocrystal composition of the present invention, adjacentdiblock polymers of a diblock polymer coating are linked together bybridging molecules. Preferably, the bridging molecules are multidentatebridging molecules having one or more reactive functional groups thatcan react with and bond to one or more hydrophilic functional groups ofthe hydrophilic end thereby crosslinking adjacent diblock polymerstogether. Therefore, the self-assembled diblock polymer coating is knittogether to form a cohesive coating around a surface-coatedsemiconductor nanocrystal composition of the present invention that willnot dissociate in water over long periods. The multidentate bridgingmolecule may have one or more than one type of reactive functionalgroup. Non-limiting examples of such reactive functional groups includehydroxy (OH), carboxylate (COOH), amine (NH.sub.2) groups, and anycombinations thereof. In a preferred embodiment, a bridging molecule isdiamine, 2,2′-(ethylenedioxy) bis(ethylamine) and the amine functionalgroups on the diamine react with hydrophilic functional groups that arecarboxylate groups on a hydrophilic end of a diblock polymer to form astable peptide bond.

In another embodiment, the water-stabilizing layer is a layer ofPEGylated phospholipids. In a preferred embodiment, the hydrophobic endof the PEGylated phospholipid comprises fatty acyl groups in aphospholipid such as 1,2-di(fatty acyl)-sn-glycero-3-phosphoethanolamineand the hydrophilic end is polyethylene glycol.

The performance of semiconductor nanocrystal complexes of the presentinvention as dependable biological research tools is related to theirability to withstand the stringent conditions found in most cellularcontexts. Oxidative stress, changes in salt concentration, pH, andtemperature, as well as proteolytic susceptibility are some examples ofthe conditions these nanocrystals need to withstand in order to beuseful in aqueous biological assays.

In addition to providing a layer of water stability to the semiconductornanocrystal composition, water-stabilizing layer can also providesurface exposed functional groups to facilitate the conjugation ofligands or tertiary molecules for target specific applications. This isthe layer of the semiconductor nanocrystal complex that opensopportunity for the biologist to exploit an entire host of molecularinteractions. For example, the semiconductor nanocrystal surfaces can be‘tagged’ with a bio-recognition molecule (e.g., antibody, peptide, smallmolecule drug or nucleic acid) designed to target only the molecularsignature of interest (e.g., cell surface receptor proteins, viral DNAsequences, disease antigens.) The interaction of the ‘tagged’semiconductor nanocrystal with its target could then be visualized withthe appropriate fluorescence detection and imaging equipment.

Functional groups exposed on the surface of a water-stable semiconductornanocrystal complex can be coupled to nucleic acids, proteins,antibodies, or small molecules and can serve as the basis for many typesof in vitro detection assays. Some examples of applicable assaysinclude, DNA/RNA assays and microarrays; high throughput screens; wholeblood and tissue screening in medical diagnostics; immunoassays, dotblots and other membrane-based detection technologies. Functional groupsinclude but are not limited to alcohol (OH), carboxylate (COOH), andamine (NH2), hydroxy, carboxyl, sulfonates, phosphates, nitrates, andany combinations thereof. More than one type of functional group may bepresent on the surface of the water-stabilizing layer. In addition tocomprising functional groups on its surface, a water-stablesemiconductor nanocrystal complex may comprise one or more tertiarymolecules.

The term tertiary molecule refers to any molecule that can be covalentlycoupled to the water-stable semiconductor nanocrystal complex. Thecoupling of tertiary molecules to a water-stable semiconductornanocrystal complex is achieved by reacting functional groups present onthe tertiary molecule with hydrophilic functional groups present on thewater-stabilizing layer of a water-stable semiconductor nanocrystalcomplex. Tertiary molecules include members of specific binding pairssuch as, for example, an antibody, antigen, hapten, antihapten, biotin,avidin, streptavidin, IgG, protein A, protein G, drug receptor, drug,toxin receptor, toxin, carbohydrate, lectin, peptide receptor, peptide,protein receptor, protein, carbohydrate receptor, carbohydrate,polynucleotide binding protein, polynucleotide, DNA, RNA, aDNA, aRNA,enzyme, and substrate. Other non-limiting examples of tertiary moleculesinclude a polypeptide, glycopeptide, peptide nucleic acid,oligonucleotide, aptamer, cellular receptor molecule, enzyme cofactor,oligosaccharide, a liposaccharide, a glycolipid, a polymer, a metallicsurface, a metallic particle, and a organic dye molecule. Depending onthe material used for the water-stabilizing layer, the tertiary moleculemay be part of the water-stabilizing layer. For example, in the eventthat the water-stabilizing layer comprises a biotin terminated lipid,then the tertiary molecule (the biotin) would be a part of thewater-stabilizing layer. One or more of the same or different tertiarymolecules can be coupled to a water-stable semiconductor nanocrystalcomplex of the present invention.

An example conjugation method to conjugate tertiary molecules to thefunctional groups of a water-stabilizing layer of a water-stablesemiconductor nanocrystal complex is illustrated in FIG. 4. Using thisprotocol, many semiconductor nanocrystals complexes that possessfunctional moieties suitable for conjugation to biomolecules can beprepared. The functional groups to be employed include, but are notlimited to, carboxylic acids, amines, sulfhydryls and maleimides.Established protein or nucleic acid conjugation protocols can then beused to generate customized ligand-bound semiconductor nanocrystals in astraightforward manner. The method depicted in FIG. 4, uses EDC(1-ethyl-3(3-dimethylaminopropyl) carbodimide HCl) and sulfo-NHS(N-hydrocylsulfo-succinimide) to form active esters on the surface ofthe semiconductor nanocrystal complex. Once the unreacted EDC andsulfo-NHS are removed, a protein of interest may be added andefficiently conjugated to a water-stable semiconductor nanocrystalcomplex.

The present invention also provides a substantially monodispersepopulation of water-stable semiconductor nanocrystal complexes, asdescribed above.

FIG. 5 provides an exemplary method of making a water-stable nanocrystalcomplex of the present invention. Although the exemplary method will bedescribed with respect to the preparation of a InGaP semiconductornanocrystal core, a metal layer of Zn and a ZnS shell, it will beappreciated that other types and combinations of semiconductor cores,metal layers, and semiconductor shells may be used.

In step 310, a semiconductor nanocrystal core is prepared. There arenumerous ways to prepare semiconductor nanocrystal cores some of whichhave been described above. The specific procedure for growing a coresemiconductor nanocrystal comprising a III-V semiconductor material isdescribed below. Although, the procedure is described with respect tothe preparation of an InGaP core, a ternary semiconductor, a binarysemiconductor material may be grown using the teachings of theprocedure. One method of preparing a semiconductor nanocrystal coreinvolves InGaP in TOPO. In order to isolate the semiconductornanocrystals cores from a TOPO solution, methanol and toluene can beadded to the solution, as semiconductor nanocrystal cores prepared inTOPO are insoluble in methanol and soluble in toluene. Enough methanolshould be added to precipitate out the semiconductor nanocrystal coresand enough toluene should be added to ensure not to precipitate any ofthe radical complexes created during the preparation of thesemiconductor nanocrystal cores. The result of washing the semiconductornanocrystal cores in methanol and toluene results in semiconductornanocrystals cores precipitating out of solution.

In step 320, the metal layer is added to the semiconductor nanocrystalcore to encapsulate the semiconductor nanocrystal core (or to otherwiseform a metal layer on the outer surface of the semiconductor nanocrystalcore). Zinc oleate in TOP dissolved in methanol, for example, can beused as precursors to create the metal layer, although many otherprecursors may be used. For example, diethylzinc, zinc acetate, zincoxalate, zinc stearate, and zinc oxide can be used as precursors for thepreparation of a metal layer of Zn. It is also appreciated that manyother precursors exist and may be used for the preparation of a metallayer comprising metals other than Zn.

Once the solution of a metal precursor(s) is prepared, it is added tothe semiconductor nanocrystal core solution resulting in a solution ofsemiconductor nanocrystal cores, purified solvent, and metal precursor.This resulting solution can then be heated and remain heated until themetal layer is formed on the semiconductor nanocrystal core. In theprocedure described below, a microwave synthesis machine is used for thereaction. It should be appreciated that in the event that traditionalheating methods are used for the reaction, the amount of time necessaryto form the metal layer as well as the temperature at which the reactionbest occurs may vary. The resulting mixture includes semiconductornanocrystal cores with a metal layer and the ligands used to prepare thereaction.

In step 330, the semiconductor nanocrystal cores with the metal layerare isolated. First, they can be drawn out of solution using thetechnique described in step 310. Specifically, methanol and toluene canbe added to the solution in amounts that make the nanocrystalcomposition, in this case the semiconductor nanocrystal core and themetal layer, insoluble. The compositions will precipitate out of thesolution resulting in a semiconductor nanocrystal composition comprisinga semiconductor nanocrystal core having an outer surface and a metallayer attached to the outer surface.

Alternatively, once the compositions comprising the semiconductornanocrystal core and the metal layer are drawn out of the solution, ashell can be added using known shelling techniques. Such an alternativeembodiment involves step 340—preparing the semiconductor nanocrystalcore with metal layer formed thereon for shelling. In one exemplaryembodiment, ZnS is grown on the core semiconductor nanocrystal with ametal layer. Initially, the isolated nanocrystal core of the III-Vsemiconductor nanocrystal material and the metal layer can be dissolvedin toluene and the resulting solution can be introduced into a solutioncontaining a ligand. The ligand can be, for example, technical gradeTOPO or TOP that has been purified using the technique described in step320. The toluene can then be removed leaving a solution of the III-Vcore nanocrystals with a metal layer of Zn.

In step 350, the shell is added to the semiconductor nanocrystal corehaving a metal layer thereon. To form the shell, two precursor solutionscan be added to the solution of the semiconductor nanocrystal core withmetal layer thereon. The first solution can be a metal precursor in aligand, the second solution can be (TMS)₂S in TOP. Enough of thesolutions is added to allow ZnS to shell the surface of thesemiconductor nanocrystal core with metal layer thereon.

An alternative to shelling the semiconductor nanocrystal that can resultin an outer coating of a desired semiconductor material is through theaddition of a chalcagonide, pnictide or a non-metal anion layer and theaddition of a second metal layer. Treating a semiconductor nanocrystalwith an anion and then adding a second metal layer stabilizes theluminescent properties of the resulting composition. Additionally,treating the nanocrystal with an anion and an additional metal layercreates a resulting composition that can be stably added to a variety ofsolvents including water and other organic and inorganic solvents withlittle or no loss of fluorescence using standard nanocrystalchemistries. The procedure for adding the non-metal anion layer and thesecond metal layer is described below in detail.

One or more anions may be attached to the semiconductor nanocrystal corecontaining the newly added metal layer. Anion layer may include anynumber, type, combination, and arrangement of anions. For example, anionlayer may be simply a monolayer of anions added to the metal layer.Non-limiting examples of elements that may comprise the anion layerinclude group IV, V, and VI elements. As described below, after theformation of the anion layer, a metal layer maybe added to the surfaceof the anion layer.

Once the nanocrystal synthesis is complete, the desired nanocrystalcompositions precipitate out of solution. All the above described stepsmay take place under nitrogen to improve the quantum yield of theresulting nanocrystal composition. The resulting nanocrystalcompositions are more resistant to oxidation and have an increasedquantum yield over semiconductor nanocrystals of III-V made by knowntechniques. The nanocrystals resulting from step 350 may be sizeseparated. Size separating the nanocrystal yields solutions withcrystals that are monodisperse and retain the quantum yield.

In step 360 and step 370, a semiconductor nanocrystal composition ismade water-stable through the addition of a surface layer (step 360) anda water-stabilizing layer (step 370). As described above, surface layersare typically organic molecules that have a moiety with an affinity forthe surface of the nanocrystals and another moiety with an affinity fora hydrophobic solvent. Lyophilic molecules, such as TOPO (trioctylphosphine oxide), TOP (trioctyl phosphine), and TBP (tributyl phosphine)are typically used in the synthesis of nanocrystals and can remain onthe surface after preparation of the semiconductor nanocrystals or maybe added or replaced by other surfactants after synthesis. Thesurfactants tend to assemble into a coating around the nanocrystal andenable it to suspend in a hydrophobic solvent. Typically, a surfacelayer of organic molecules will be present on the surface of thenanocrystal after the completion of step 350. However, in the event thata different surface molecule is desired exchange reactions may be doneto alter the surface layer.

Water stabilizing layers are generally but not exclusively linear chainsand have a hydrophilic end section and a hydrophobic end section. Asdescribed above, this layer may comprise lipids or polymer-based smallmolecules including amino acids with a hydrophilic end section and ahydrophobic end section. Stabilizing the surface for water solubleapplications can be achieved by layering a coat of hydrophilic materialonto the semiconductor nanocrystal. The composition of this material canvary and can be selected based on the application to be used.

The above-described technique is only exemplary and other modificationsmay be made that result in semiconductor nanocrystal compositionsaccording to the present invention.

EXAMPLE 1 Preparing a III-V Semiconductor Nanocrystal Composition

The present example discloses how to prepare a stable, high luminescentquantum yield semiconductor nanocrystal composition comprising an InGaPsemiconductor nanocrystal core and a Zn metal layer formed on the outersurface of the semiconductor nanocrystal core after synthesis of thesemiconductor nanocrystal core.

First, an indium precursor is prepared. In a reaction flask, 292 g/molof 0.9 Molar indium (III) acetate (99.99% pure), is added to 367 g/molof 0.1 Molar gallium acetylacetonoate (99% pure) and 256.4 g/mol of 3Molar almitic acid (99% pure). The ingredients are mixed and heated to130° C. for 2 hours under vacuum. The vacuum removes the acetic acidthat could form. The resulting solution should be clear but may have ayellow tint. Next, a phosphorous precursor is prepared by adding 0.3 mLTMSphosphine and 4.0 mL TOP under nitrogen.

Then a zinc oleate precursor is prepared by heating at 140° C., 1.83 gof 10 mmol zinc acetate, 6.49 mL of oleic acid, and 3.5 mL of octadecenein a reaction flask for 2.5 hours. The resulting solution should beclear to yellow.

After the preparation of the precursors, the InGaP semiconductornanocrystal core is prepared.

For a InGaP semiconductor nanocrystal core with a peak emissionwavelength between 610 nm-630 nm the following procedure is followed.

In a closed reaction flask, 700 mg of the indium precursor, preparedabove is added to 0.25 g of dodecyl amine, octadecyl amine orhexadecylamine, 0.25 g of 99% pure TOPO, and 2.5 mL of toluene oroctadecene. The system is purged with nitrogen and the reaction isheated to 130° C. in a Discovery microwave system (CEM Corporation) and2.5 mL of the phosphorous precursor, prepared above, is injected intothe reaction flask and the reaction is heated to 270° C. for 8 minutes.During the heating process the reaction is cooled by an air strain inorder retain microwave flux at 300 Watts of microwave power. The flowrate of the air is adjusted to maintain the desired microwave power.Finally, the reaction is allowed to cool to 50° C. The resultingsemiconductor complex is a InGaP core semiconductor nanocrystal.

For a InGaP semiconductor nanocrystal core with a peak emissionwavelength between 630 nm-660 nm the following procedure is followed.

In a closed reaction flask, 700 mg of the indium precursor, preparedabove is added to 0.5 g of dodecyl amine, octadecyl amine orhexadecylamine, 0.5 g of 99% pure TOPO, and 2 mL of toluene oroctadecene. The system is purged with nitrogen and the reaction isheated to 140° C.-150° C. in CEM Corporation's, Discovery microwavesystem and 2.5 mL of the phosphorous precursor, prepared above, isinjected into the reaction flask and the reaction is heated to 270° C.for 8 minutes. During the heating process the reaction is cooled by anair strain in order retain microwave flux at 300 Watts of microwavepower. The flow rate of the air is adjusted to maintain the desiredmicrowave power. Finally, the reaction is allowed to cool to 50° C. Theresulting semiconductor core is a InGaP core semiconductor nanocrystal.

For a InGaP semiconductor nanocrystal core with a peak emissionwavelength between 660 nm-690 nm the following procedure is followed.

In a closed reaction flask, 700 mg of the indium precursor, preparedabove is added to 0.5 g of dodecyl amine, octadecyl amine orhexadecylamine, 0.5 g of 99% pure TOPO, and 2 mL of toluene oroctadecene. The system is purged with nitrogen and the reaction isheated to 160° C.-170° C. in CEM Corporation's, Discovery microwavesystem and 2.5 mL of the phosphorous precursor, prepared above, isinjected into the reaction flask and the reaction is heated to 270° C.for 8 minutes. During the heating process the reaction is cooled by anair strain in order retain microwave flux at 300 Watts of microwavepower. The flow rate of the air is adjusted to maintain the desiredmicrowave power. Finally, the reaction is allowed to cool to 50° C. Theresulting semiconductor core is a InGaP core semiconductor nanocrystal.

For a InGaP semiconductor nanocrystal core with a peak emissionwavelength between 690 nm-740 nm the following procedure is followed.

In a closed reaction flask, 700 mg of the indium precursor, preparedabove is added to 2 g of dodecyl amine, octadecyl amine orhexadecylamine, 2 g of 99% pure TOPO. The system is purged with nitrogenand the reaction is heated to 180° C. in CEM Corporation's, Discoverymicrowave system and 2.5 mL of the phosphorous precursor, preparedabove, is injected into the reaction flask and the reaction is heated to270° C. for 8 minutes. During the heating process the reaction is cooledby an air strain in order retain microwave flux at 300 Watts ofmicrowave power. The flow rate of the air is adjusted to maintain thedesired microwave power. Finally, the reaction is allowed to cool to 50°C. The resulting semiconductor core is a InGaP core semiconductornanocrystal.

To any of the flasks containing the InGaP semiconductor nanocrystalcomposition, the 0.8 g of the zinc oleate precursor and 0.4 ml of anoleyl alcohol is added. The oleyl alcohol allows the zinc oleate to bemore reactive and thus provides better surface converage around the coresemiconductor nanocrystal. The system is purged with nitrogen and heatedto 270° C. for 5 minutes and the reaction is allowed to cool to 50° C.During the heating process, the reaction is cooled by an air strain inorder retain microwave flux at 300 Watts of microwave power. The flowrate of the air is adjusted to maintain the desired microwave power.

After the addition of the metals to the outer surface of thesemiconductor nanocrystal core, the resulting semiconductor nanocrystalcomposition is washed. To wash the semiconductor nanocrystalcomposition, butanol and methanol are added to the reaction flask. Thesupernatants are then removed from the reaction flask by centrifugingthe reaction flask at 2000 rpm for 4 minutes and decanting thesupernatant. Once the supernatant is removed, the nanocrystalcompositions are re-solvated in an organic solvent, such as toluene.

Once solvated, the nanocrystal compositions may be treated with an anionand a second metal layer. A sulfur source may be prepared by dissolving70 mg of elemental sulfur (1 mM) in 4 mL TOP by heating in hot bath andsonication. To add this outer coating onto the nanocrystal composition,0.1 ml of hexamethyldisilathiane is dissolved in 0.9 g of TOP. 1.0 mL ofthe prepared sulfur source is added to 1.0 mL of one of the nanocrystalcomposition/solvent solutions prepared above. Then 0.8 mg of TOPO, (99%pure) is added to the flask and the system is purged with nitrogen. Themixture is heated to 70° C. for 5 minute.

The resulting solution is illuminated under UV excitation for 30 minutesor gently heated for 30 minutes. This anneals the surface because itallows the the nanocrystal surface become more reactive.

A second metal layer is next added to the semiconductor nanocrystalcomposition. To add a second metal layer onto the nanocrystalcomposition, 0.86 mg of TOP (90% pure) is added to the flask containingthe nanocrystal composition/organic solvent solution resulting from thewashing step. 0.25 g zinc oleate is then added to the flask. In order toisolate the semiconductor nanocrystal compositions, the washing step maybe performed for a third time.

EXAMPLE 2 Preparing a Water-Stable III-V Semiconductor NanocrystalComplex

The below described procedures may be used for the development ofvarious water-stable semiconductor nanocrystal complexes. Althoughspecific amounts and temperatures are given in the below describedprocedures, it is appreciated that these amounts may be varied. Thestarting material for the below described procedure are thesemiconductor nanocrystal compositions prepared and described above.

The below described procedure can be used to produce a water-stablesemiconductor nanocrystal complex comprising approximately 3 functionalgroups. Depending on the size of the semiconductor nanocrystal, theratio of lipids described below may be varied to get the appropriatenumber of functional groups.

EXAMPLE 2a Carboxy Terminated Water Stable III-V SemiconductorNanocrystal Complexes

Solution 1. 2.2 mg of any of the semiconductor nanocrystal compositionsdescribed above in toluene solution are loaded into a 15 ml centrifugetube, and 10 ml methanol is added. The solution is mixed, andcentrifuged at 4000 rpm for 3 minutes. The supernatant is removedcarefully and 3 ml of hexane is added to re-dissolve the pellet of thenanocrystals. Then 9 ml methanol is added in the tube to precipitatedown the nanocrystals again. The hexane and methanol purification stepsare repeated one more time. The precipitate pellet is dried, and thenre-dissolved into 1 ml chloroform.

Solution 2. 10 mg DSPE-PEG(2000)CarboxylicAcid(1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine-N-[Carboxy(PolyethyleneGlycol)2000] (Ammonium Salt)) and 60 mg mPEG2000PE([1,2-diacyl-sn-glycero-3-phosphoethanolamine-n-methoxy(polyethyleneglycol)-2000] (Ammonium Salt)) lipids are dissolved in 3 ml chloroformsolution. It has been found that a 1/6 ratio of carboxy lipid toammonium salt lipid allows for an optimal number of functional groups,approximately three. However, these ratios may be varied depending onthe number of functional groups desired on the semiconductor nanocrystalcomplex.

Solution 1 and solution 2 are mixed together in a 20 ml vial and theresultant solution dried under N₂. The vial is rotated slowly to make athin film on the wall while the solution is drying. The vial is heatedat 75° C. in a water bath for 2 minutes. To the heated vial, 5 mldeionized water, which has been preheated to 75° C., is added. Then thevial is capped, and the solution is vortexed until all the nanocrystalsare dissolved. Then the solution is sonicated for 1 minute.

The solution is transferred into a 15 ml centrifuge tube, centrifuged at4000 rpm for 5 minutes. The clear supernatant is loaded into a 10 mlsyringe, and is filtrated through a 0.2 μm filter. Afterwards, thefiltrated solution is loaded into two 11 ml ultracentrifuge tubes andcentrifuged at 65000 rpm for 1 hour. The supernatant is removedcarefully, and the precipitates is re-dissolved into deionized water.The ultracentrifuge purification step is repeated one more time. Thepellets are reconstituted into 4 ml deionized water and stored at 4° C.

EXAMPLE 2b Amine Terminated Water-Stable III-V Semiconductor NanocrystalComplexes

Solution 1. 2.2 mg of any of the semiconductor nanocrystal compositionsdescribed above is loaded into a 15 ml centrifuge tube, and 10 mlmethanol is added. The solution is mixed, and centrifuged at 4000 rpmfor 3 minutes. The supernatant is removed carefully; and 3 ml of hexaneis added to re-dissolve the pellet of the nanocrystals. Then 10 mlmethanol is added in the tube to precipitate down the nanocrystalsagain. The hexane and methanol purification steps are repeated one moretime. The precipitate pellet is dried and then re-dissolved in 1 mlchloroform.

Solution 2. 10 mg DSPE-PEG(2000)Amine(1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine-N-[Amino(PolyethyleneGlycol)2000] (Ammonium Salt)) and 60 mg mPEG2000PE([1,2-diacyl-sn-glycero-3-phosphoethanolamine-n-methoxy(polyethyleneglycol)-2000] (Ammonium Salt)) lipids are dissolved in 3 ml chloroformsolution. It has been found that a 1/6 ratio of amine lipid to ammoniumsalt lipid allows for an optimal number of functional groups. However,these ratios may be varied depending on the number of functional groupsdesired on the semiconductor nanocrystal complex. Solution 1 andsolution 2 are mixed together in a 20 ml vial and the resultant solutionwas dried under N₂. The vial is rotated slowly to make a thin film onthe wall while the solution is drying. The vial is heated at 75° C. in awater bath for 2 minutes. To the heated vial, 5 ml deionized water,which had been preheated to 75° C., is added. Then the vial is capped,and the solution is vortexed until all the nanocrystals are dissolved.Then the solution is sonicated for 1 minute.

The solution is transferred into a 15 ml centrifuge tube, centrifuged at4000 rpm for 5 minutes. The clear supernatant is loaded into a 10 mlsyringe, and is filtrated through a 0.2 μm filter. Afterwards, thefiltrated solution is loaded into two 11 ml ultracentrifuge tubes,centrifuged at 65000 rpm for 1 hour. The supernatant is removedcarefully, and re-dissolved into deionized water. The ultracentrifugepurification step is repeated one more time and the pellets arereconstituted into 4 ml deionized water and stored at 4° C.

EXAMPLE 2c Biotin Terminated Water-Stable III-V SemiconductorNanocrystal Complexes

Solution 1. 2.2 mg of any of the semiconductor nanocrystal compositionsdescribed above is loaded into a 15 ml centrifuge tube, and 10 mlmethanol is added. The solution is mixed, and centrifuged at 4000 rpmfor 3 minutes. The supernatant is removed and 3 ml of hexane is added tore-dissolve the pellet of the nanocrystals. Then 9 ml methanol is addedinto the tube to precipitate down the nanocrystals again. The hexane andmethanol purification step are repeated one more time. The precipitatepellet is dried under air, and then re-dissolved into 1 ml chloroform.

Solution 2. 10 mg DPPE-PEG(2000) biotinlipids(1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine-N-[Biotinyl(PolyethyleneGlycol)2000 (Ammonium Salt)) and 60 mg mPEG2000PE([1,2-diacyl-sn-glycero-3-phosphoethanolamine-n-methoxy(polyethyleneglycol)-2000] (Ammonium Salt)) lipids are dissolved in 3 ml chloroformsolution. It has been found that a 1/6 ratio of biotin lipid to ammoniumsalt lipid allows for an optimal number of functional groups,approximately three in this example. However, these ratios may be varieddepending on the number of functional groups desired on thesemiconductor nanocrystal complex.

Solution 1 and solution 2 were mixed together in a 20 ml vial and theresultant solution are dried under N₂. The vial was rotated slowly tomake a thin film on the wall while the solution is drying. The vial isheated at 75° C. in a water bath for 2 minutes. To the heated vial, 5 mldeionized water which has been preheated to 75° C. is added. Then thevial is capped, and the solution is vortexed until all the nanocrystalswere dissolved. Then the solution is sonicated for 1 minute.

The solution is transferred into a 15 ml centrifuge tube, centrifuged at4000 rpm for 5 minutes. The clear supernatant is loaded into a 10 mlsyringe, and is filtrated through a 0.2 μm filter. Afterwards, thefiltrated solution is loaded into two 11 ml ultracentrifuge tubes,centrifuged at 65,000 rpm for 1 hour. The supernatant is removedcarefully, and the precipitates are re-dissolved into deionized water.The ultracentrifuge purification step is repeated one more time. Thepellets are reconstitute into 4 ml deionized water and stored at 4° C.

The foregoing description and example have been set forth merely toillustrate the invention and are not intended as being limiting. Each ofthe disclosed aspects and embodiments of the present invention may beconsidered individually or in combination with other aspects,embodiments, and variations of the invention. In addition, unlessotherwise specified, none of the steps of the methods of the presentinvention are confined to any particular order of performance.Modifications of the disclosed embodiments incorporating the spirit andsubstance of the invention may occur to persons skilled in the art andsuch modifications are within the scope of the present invention.Furthermore, all references cited herein are incorporated by referencein their entirety.

1. A water-stable semiconductor nanocrystal complex comprising: asemiconductor nanocrystal composition comprising: a III-V semiconductornanocrystal core; and a surface layer comprising molecules having amoiety with an affinity for the semiconductor nanocrystal compositionand a moiety with an affinity for a hydrophobic solvent; and awater-stabilizing layer comprising a diblock polymer that surrounds thesemiconductor nanocrystal composition to provide a micelle and having ahydrophobic end for interacting with the surface layer and a hydrophilicend, wherein the water-stable semiconductor nanocrystal complex iselectronically and chemically stable with a luminescent quantum yield ofat least 45% as measured under ambient conditions.
 2. The water-stablesemiconductor nanocrystal complex of claim 1, wherein the semiconductornanocrystal composition comprises: a metal layer formed on an outersurface of the semiconductor nanocrystal core; and a shell comprising asemiconductor material overcoating the metal layer.
 3. The water-stablesemiconductor nanocrystal complex of claim 1, wherein the semiconductornanocrystal composition comprises: a metal layer formed on an outersurface of the semiconductor nanocrystal core; and a shell comprising ananion layer overcoating the metal layer and a second metal layerovercoating the anion layer.
 4. The water-stable semiconductornanocrystal complex of claim 1, wherein the semiconductor nanocrystalcore comprises InP.
 5. The water-stable semiconductor nanocrystalcomplex of claim 1, wherein the semiconductor nanocrystal core is aternary semiconductor nanocrystal.
 6. The water-stable semiconductornanocrystal complex of claim 5, wherein the ternary semiconductornanocrystal is InGaP.
 7. The water-stable semiconductor nanocrystalcomplex of claim 1, wherein the luminescent quantum yield is at least55% as measured under ambient conditions.
 8. The water-stablesemiconductor nanocrystal complex of claim 2, wherein the semiconductornanocrystal complex comprises InP, the shell comprises ZnS, and themetal layer comprises Zn.
 9. The water-stable semiconductor nanocrystalcomplex of claim 1, wherein the diblock polymer comprises one selectedfrom poly(acrylic acid-b-methyl methacrylate), poly(methylmethacrylate-b-sodium acrylate), poly(t-butyl methacrylate-b-ethyleneoxide), poly(methyl methacrylate-b-sodium methacrylate), poly (methylmethacrylate-b-N-methyl 1-4vinyl pyridinium iodide), poly(methylmethacrylate-b-N,N-dimethyl acrylamide), poly(butadiene-b-methacrylateacid and sodium salt), poly(butadiene(1,2 addition)-b-acrylic acid),poly(butadiene(1,2 addition)-b-sodium acrylate), poly(butadiene(1,4addition)-b-acrylic acid), poly(butadiene(1,4 addition)-b-sodiumacrylate), poly(butadiene(1,4 addition)-b-ethylene oxide),poly(butadiene(1,2 addition)-b-ethylene oxide), poly(styrene-b-acrylicacid), poly(styrene-b-acrylamide), poly(styrene-b-cesium acrylate),poly(styrene-b-sodium acrylate), poly(styrene-b-ethylene oxide),poly(styrene-b-methacrylic acid), poly(styrene-b-sodium methacrylate),and a combination thereof.
 10. The water-stable semiconductornanocrystal complex of claim 1, wherein the hydrophilic end of thewater-stabilizing layer comprises a functional group for coupling one ormore tertiary molecule.
 11. The water-stable semiconductor nanocrystalcomplex of claim 10, further comprising one or more tertiary moleculecoupled to the functional group.
 12. The water-stable semiconductornanocrystal complex of claim 11, wherein the one or more tertiarymolecule is a member of a specific binding pair.
 13. The water-stablesemiconductor nanocrystal complex of 11, wherein the member of thespecific binding pair is selected from the group consisting of antibody,antigen, hapten, antihapten, biotin, avidin, streptavidin, IgG, proteinA, protein G, drug receptor, drug, toxin receptor, toxin, carbohydrate,lectin, peptide receptor, peptide, protein receptor, protein,carbohydrate receptor, carbohydrate, polynucleotide binding protein,polynucleotide, DNA, RNA, aDNA, aRNA, enzyme, substrate.
 14. Asubstantially monodisperse population of the water-stable semiconductornanocrystal complexes of claim
 1. 15. A method of making a water-stablesemiconductor nanocrystal complex comprising: synthesizing asemiconductor nanocrystal core comprising a III-V semiconductornanocrystal material; forming a surface layer either directly orindirectly on the semiconductor nanocrystal core; and forming awater-stabilizing layer on the surface layer, the water-stabilizinglayer having a hydrophobic end for interacting with the surface layerand a hydrophilic end, wherein the water-stable semiconductornanocrystal complex is electronically and chemically stable with aluminescent quantum yield of at least 45% as measured under ambientconditions.
 16. The method of claim 15, further comprising forming ametal layer on the semiconductor nanocrystal core.
 17. The method ofclaim 16, further comprising overcoating the metal layer with an anionlayer.
 18. The method of claim 17, further comprising overcoating theanion layer with a second metal layer.
 19. The method of claim 15,further comprising overcoating the metal layer with a shell comprising asemiconductor material.